Formation of intracellular vesicles within the Gram+ Lactococcus lactis induced by the overexpression of Caveolin-1β

International audienceBackground: Caveolae are invaginated plasma membrane domains of 50-100 nm in diameter involved in many important physiological functions in eukaryotic cells. They are composed of different proteins, including the membrane-embedded caveolins and the peripheric cavins. Caveolin-1 has already been expressed in various expression systems (E. coli, insect cells, Toxoplasma gondii, cell-free system), generating intracellular caveolin-enriched vesicles in E. coli, insect cells and T. gondii. These systems helped to understand the protein insertion within the membrane and its oligomerization. There is still need for fundamental insights into the formation of specific domains on membrane, the deformation of a biological membrane driven by caveolin-1, the organization of a caveolar coat, and the requirement of specific lipids and proteins during the process. The aim of this study was to test whether the heterologously expressed caveolin-1β was able to induce the formation of intracellular vesicles within a Gram + bacterium, Lactococcus lactis, since it displays a specific lipid composition different from E. coli and appears to emerge as a good alternative to E. coli for efficient overexpression of various membrane proteins. Results: Recombinant bacteria transformed with the plasmid pNZ-HTC coding for the canine isoform of caveolin-1β were shown to produce caveolin-1β, in its functional oligomeric form, at a high expression level unexpected for an eukaryotic membrane protein. Electron microscopy revealed several intracellular vesicles from 30 to 60 nm, a size comparable to E. coli h-caveolae, beneath the plasma membrane of the overexpressing bacteria, showing that caveolin-1β is sufficient to induce membrane vesiculation. Immunolabelling studies showed antibodies on such neo-formed intracellular vesicles, but none on plasma membrane. Density gradient fractionation allowed the correlation between detection of oligomers on Western blot and appearance of vesicles measurable by DLS, showing the requirement of caveolin-1β oligomerization for vesicle formation. Conclusions: Lactococcus lactis cells can heterologously overexpress caveolin-1β, generating caveolin-1β enriched intracellular neo-formed vesicles. These vesicles might be useful for potential co-expression of membrane proteins of pharmaceutical interest for their simplified functional characterization

Cavezzo, the first Italian meteorite recovered by the PRISMA fireball network. Orbit, trajectory, and strewn-field

International audienceTwo meteorite pieces have been recovered in Italy, near the town of Cavezzo (Modena), on 2020 January 4th. The associated fireball was observed on the evening of New Year's Day 2020 by eight all-sky cameras of the PRISMA fireball network, a partner of FRIPON. The computed trajectory had an inclination angle of approximately 68° and a velocity at infinity of 12.8 km s-1. Together with the relatively low terminal height, estimated as 21.5 km, those values were indicating the significant possibility of a meteorite dropping event, as additionally confirmed by the non-zero residual total mass. The strewn-field was computed taking into account the presence of two bright light flashes, revealing that the meteoroid had been very likely subject to fragmentation. Three days after the event, two samples, weighing 3.1 and 52.2 g, were collected as a result of a dedicated field search and thanks to the involvement of the local people. The two pieces were immediately recognized as freshly fallen fragments of meteorite. The computed orbital elements, compared with the ones of known Near-Earth Asteroids from the NEODyS database, are compatible with one asteroid only; 2013 VC10. The estimated original mass of the meteoroid, 3.5 kg, and size, approximately 13 cm, is so far the smallest among the current 35 cases in which meteorites were recovered from precise strewn-field computation thanks to observational data. This result demonstrates the effectiveness of accurate processing of fireball network data even on challenging events generated by small size meteoroids

FRIPON: a worldwide network to track incoming meteoroids

(IF 5.80; Q1)International audienceContext. Until recently, camera networks designed for monitoring fireballs worldwide were not fully automated, implying that in case of a meteorite fall, the recovery campaign was rarely immediate. This was an important limiting factor as the most fragile-hence precious-meteorites must be recovered rapidly to avoid their alteration. Aims. The Fireball Recovery and InterPlanetary Observation Network (FRIPON) scientific project was designed to overcome this limitation. This network comprises a fully automated camera and radio network deployed over a significant fraction of western Europe and a small fraction of Canada. As of today, it consists of 150 cameras and 25 European radio receivers and covers an area of about 1.5 × 10 6 km 2. Methods. The FRIPON network, fully operational since 2018, has been monitoring meteoroid entries since 2016, thereby allowing the characterization of their dynamical and physical properties. In addition, the level of automation of the network makes it possible to trigger a meteorite recovery campaign only a few hours after it reaches the surface of the Earth. Recovery campaigns are only organized for meteorites with final masses estimated of at least 500 g, which is about one event per year in France. No recovery campaign is organized in the case of smaller final masses on the order of 50 to 100 g, which happens about three times a year; instead, the information is delivered to the local media so that it can reach the inhabitants living in the vicinity of the fall. Results. Nearly 4000 meteoroids have been detected so far and characterized by FRIPON. The distribution of their orbits appears to be bimodal, with a cometary population and a main belt population. Sporadic meteors amount to about 55% of all meteors. A first estimate of the absolute meteoroid flux (mag <-5; meteoroid size ≥∼1 cm) amounts to 1250/yr/10 6 km 2. This value is compatible with previous estimates. Finally, the first meteorite was recovered in Italy (Cavezzo, January 2020) thanks to the PRISMA network, a component of the FRIPON science project

FRIPON: A worldwide network to track incoming meteoroids

Context. Until recently, camera networks designed for monitoring fireballs worldwide were not fully automated, implying that in case of a meteorite fall, the recovery campaign was rarely immediate. This was an important limiting factor as the most fragile - hence precious - meteorites must be recovered rapidly to avoid their alteration. Aims. The Fireball Recovery and InterPlanetary Observation Network (FRIPON) scientific project was designed to overcome this limitation. This network comprises a fully automated camera and radio network deployed over a significant fraction of western Europe and a small fraction of Canada. As of today, it consists of 150 cameras and 25 European radio receivers and covers an area of about 1.5 × 106km2. Methods. The FRIPON network, fully operational since 2018, has been monitoring meteoroid entries since 2016, thereby allowing the characterization of their dynamical and physical properties. In addition, the level of automation of the network makes it possible to trigger a meteorite recovery campaign only a few hours after it reaches the surface of the Earth. Recovery campaigns are only organized for meteorites with final masses estimated of at least 500 g, which is about one event per year in France. No recovery campaign is organized in the case of smaller final masses on the order of 50 to 100 g, which happens about three times a year; instead, the information is delivered to the local media so that it can reach the inhabitants living in the vicinity of the fall. Results. Nearly 4000 meteoroids have been detected so far and characterized by FRIPON. The distribution of their orbits appears to be bimodal, with a cometary population and a main belt population. Sporadic meteors amount to about 55% of all meteors. A first estimate of the absolute meteoroid flux (mag &lt; -5; meteoroid size ≥∼1 cm) amounts to 1250/yr/106km2. This value is compatible with previous estimates. Finally, the first meteorite was recovered in Italy (Cavezzo, January 2020) thanks to the PRISMA network, a component of the FRIPON science project

FRIPON: A worldwide network to track incoming meteoroids

Context. Until recently, camera networks designed for monitoring fireballs worldwide were not fully automated, implying that in case of a meteorite fall, the recovery campaign was rarely immediate. This was an important limiting factor as the most fragile - hence precious - meteorites must be recovered rapidly to avoid their alteration. Aims. The Fireball Recovery and InterPlanetary Observation Network (FRIPON) scientific project was designed to overcome this limitation. This network comprises a fully automated camera and radio network deployed over a significant fraction of western Europe and a small fraction of Canada. As of today, it consists of 150 cameras and 25 European radio receivers and covers an area of about 1.5 × 106km2. Methods. The FRIPON network, fully operational since 2018, has been monitoring meteoroid entries since 2016, thereby allowing the characterization of their dynamical and physical properties. In addition, the level of automation of the network makes it possible to trigger a meteorite recovery campaign only a few hours after it reaches the surface of the Earth. Recovery campaigns are only organized for meteorites with final masses estimated of at least 500 g, which is about one event per year in France. No recovery campaign is organized in the case of smaller final masses on the order of 50 to 100 g, which happens about three times a year; instead, the information is delivered to the local media so that it can reach the inhabitants living in the vicinity of the fall. Results. Nearly 4000 meteoroids have been detected so far and characterized by FRIPON. The distribution of their orbits appears to be bimodal, with a cometary population and a main belt population. Sporadic meteors amount to about 55% of all meteors. A first estimate of the absolute meteoroid flux (mag < -5; meteoroid size ≥∼1 cm) amounts to 1250/yr/106km2. This value is compatible with previous estimates. Finally, the first meteorite was recovered in Italy (Cavezzo, January 2020) thanks to the PRISMA network, a component of the FRIPON science project

FRIPON: a worldwide network to track incoming meteoroids

Context: Until recently, camera networks designed for monitoring fireballs worldwide were not fully automated, implying that in case of a meteorite fall, the recovery campaign was rarely immediate. This was an important limiting factor as the most fragile – hence precious – meteorites must be recovered rapidly to avoid their alteration. Aims: The Fireball Recovery and InterPlanetary Observation Network (FRIPON) scientific project was designed to overcome this limitation. This network comprises a fully automated camera and radio network deployed over a significant fraction of western Europe and a small fraction of Canada. As of today, it consists of 150 cameras and 25 European radio receivers and covers an area of about 1.5 × 106 km2. Methods: The FRIPON network, fully operational since 2018, has been monitoring meteoroid entries since 2016, thereby allowing the characterization of their dynamical and physical properties. In addition, the level of automation of the network makes it possible to trigger a meteorite recovery campaign only a few hours after it reaches the surface of the Earth. Recovery campaigns are only organized for meteorites with final masses estimated of at least 500 g, which is about one event per year in France. No recovery campaign is organized in the case of smaller final masses on the order of 50 to 100 g, which happens about three times a year; instead, the information is delivered to the local media so that it can reach the inhabitants living in the vicinity of the fall. Results: Nearly 4000 meteoroids have been detected so far and characterized by FRIPON. The distribution of their orbits appears to be bimodal, with a cometary population and a main belt population. Sporadic meteors amount to about 55% of all meteors. A first estimate of the absolute meteoroid flux (mag < –5; meteoroid size ≥~1 cm) amounts to 1250/yr/106 km2. This value is compatible with previous estimates. Finally, the first meteorite was recovered in Italy (Cavezzo, January 2020) thanks to the PRISMA network, a component of the FRIPON science project